Preferential binding of yeast tRNA ligase to pre-tRNA substrates
Open Access
- 1 January 1991
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 19 (8) , 1853-1860
- https://doi.org/10.1093/nar/19.8.1853
Abstract
Joining of tRNA halves during splicing in extracts of Saccharomyces cerevisiae requires each of the three enzymatic activities associated with the tRNA ligase polypeptide. Joining is most efficient for tRNA as opposed to oligonucleotide substrates and is sensitive to single base changes at a distance from splice sites suggesting considerable specificity. To examine the basis for this specificity, binding of ligase to labeled RNA substrates was measured by native gel electrophoresis. Ligase bound tRNA halves with an association constant 1600-fold greater than that for a nonspecific RNA. Comparison of binding of a series of tRNA processing intermediates revealed that tRNA-structure, particularly in the region around the splice sites, contributes to specific binding. Finally, the ligase was shown to form multiple, discrete complexes with tRNA substrates. The basis for recognition by ligase and its role in a tRNA processing pathway are discussed.Keywords
This publication has 42 references indexed in Scilit:
- Effects of tRNA-intron structure on cleavage of precursor tRNAs by RNase P fromSaccharomyces cerevisiaeNucleic Acids Research, 1988
- Directed Deletion of a Yeast Transfer RNA Intervening SequenceScience, 1980
- LIGAND: A versatile computerized approach for characterization of ligand-binding systemsAnalytical Biochemistry, 1980
- Order and intracellular location of the events involved in the maturation of a spliced tRNANature, 1980
- Splicing of yeast tRNA precursors: a two-stage reactionCell, 1979
- Splicing of yeast tRNA precursors: structure of the reaction intermediatesCell, 1979
- Nucleic acid binding properties of Escherichia coli ribosomal protein S1Journal of Molecular Biology, 1978
- Role of amino acids in osmoregulation of non-halophilic bacteriaNature, 1975
- Amino-acid metabolism during osmotic stress in isolated axons of Callinectes sapidusLife Sciences, 1974
- Extension of the theory of linked functions to incorporate the effects of protein hydrationJournal of Molecular Biology, 1969